Determination of sulfamethazine and trimethoprim in liquid feed premixes by HPLC and diode array detection, with an analysis of the uncertainty of the analytical results.

Sulfamethazine (SMZ) and trimethoprim (TMP) are antibacterials used in veterinary practice. This paper describes a method for their determination in veterinary liquid feed premixes that is based on liquid chromatography with diode array detection. Gradient elution with methanol and ammonium acetate achieved excellent separation of the two analytes within 15 min without any interference from the matrix. Absorbance of the column effluent was monitored at 264 nm for SMZ and at 230 nm for TMP. Detailed analyses of the uncertainties of determinations afford estimated expanded uncertainties of, respectively, 0.2 and 0.1 w/v % for typical SMZ and TMP concentrations of 10.7 and 2.1 w/v %, respectively. At the lower end of the calibrated range of the method, the dominant source of uncertainty is the preparation of standards and the construction of the calibration line.     

Correct equations for comparing models in diallel analysis

SUMMARY: Correct equations are given to express the parameter estimates of four models for complete diallels as a function of the parameter estimates in the model of Eisen et al. (1983). In recent literature these equations have been partly incorrect. ZUSAMMENFASSUNG: Korrekte Gleichungen für den Vergleich von Modellen in der Diallelanalyse Für die Darstellung der Parametersch?tzwerte von vier Modellen für vollst?ndige Diallele als Funktion der Parametersch?tzwerte des Modells von Eisen et al. (1983), die in der Literatur teilweise fehlerhaft erfolgte, werden korrekte Formeln angegeben.     

First record of bacillary rickettsia-like organisms in European tick Dermacentor marginatus (Sulzer).

Bacillary rickettsia-like organisms (BRLO) were found in the tick Dermacentor marginatus. They are gram-negative and differ from common bacteria and reckettsiae both in the cultivation conditions and morphology. BRLO are non-pathogenic for ticks and guinea pigs. In our studies they were isolated on half-engorged females of D. marginatus, on which they are still maintained.     

Histological studies on cow intestinal mucosa and electron microscopic studies of the ruminal epithelium of cows fed a deficient diet for a prolonged time]

Histological and histochemical studies were carried out on the gastro-intestinal mucosa of three experimental cows (roughages/maize silage) and three control animals. The animals were slaughtered on termination of the long-term trial. Mucosa samples were taken, for further study, from the rumen, the duodenum, the jejunum, the large intestine and the appendix. Histochemical analysis did not reveal any essential differences in the activities of non-specific esterase, alkaline and acid phosphatase and lactic acid dehydrogenase in the mucosa of the rumen, the large and the small intestine and the appendix of both the experimental animals and the controls. The experimental animals were found to exhibit a higher rate of glutamate-dehydrogenase activity in the ruminal mucosa and in the mucosa of the large and small intestine. A higher succinate dehydrogenase activity was observed in the ruminal mucosa of the experimental animals, relative to that of the controls, while the activity in the intestinal mucosa was decreased. Only slight changes were noted in the activity of the enzymatic systems tested. Electron microscopic studies did not reveal any differences in the ultrastructure of the epithelial cells of the ruminal mucosa in both the experimental animal and the controls.     

Testing of the protective effect of cell-free lyophilized vaccine against Marek's disease in field trials]

The protective effect of a lyophilized vaccine against Marek's disease ("Keramvac"--Pfizer), prepared from a cell-free turkey's herpesvirus (strain FC 126), was compared in field trials with losses in the group of non-vaccinated chickens. Under the indicated conditions, the vaccine had only a 50.86% effectiveness. The possible causes of the reduced vaccination effect are discussed with regard to the pathomorphological and virological findings suggesting, among others, an increased incidence of the symptoms of the classical form of Marek's disease in the population investigated.     

Pregnancy Rates in Repeat-breeder Heifers Following Multiple Artificial Inseminations during Spontaneous Oestrus

Hormonal asynchronies during oestrus, related to the presence of suprabasal plasma-progesterone (P4) concentrations and a delayed ovulation, interfere with the fertility of repeat-breeder heifers (RBH). Since tubal dysfunction can occur in connection with hormonal asynchronies and constrained availability of fertile spermatozoa at the time of ovulation, the present study tested the hypothesis that frequent sperm deposition from onset of oestrus to ovulation may improve pregnancy rates in RBH. Five RBH and five virgin heifers (VH; controls) were repeatedly artificially inseminated (AI) at 6 h intervals from onset of oestrus to spontaneous ovulation. Hormone analyses revealed suprabasal P₄ concentrations and a delay in the occurrence of the luteinising hormone (LH) surge, but a normal cortisol profile in RBH. Compared with controls, RBH presented longer interval from onset of oestrus to ovulation, and therefore, received more AIs. Pregnancy rates in RBH reached control levels (60%; NS), indicating that the hypothesis might be correct. Pregnancy rates in VH were below the expected range, presumably attributed to a deleterious influence of the frequent handling. The study suggests that pregnancy rates can be improved in RBH by frequent AI in relation to spontaneous ovulation. However, this practice of repeated manipulations, while seeming not to show adverse effects, lacks practicality for routine use.     

C-reactive protein measurement in canine saliva.

An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.     

Detection of the Defoliating Pathotype of Verticillium dahliae in Infected Olive Plants by Nested PCR

Spread of Verticillium wilt into newly established olive orchards in Andalucía, southern Spain, has caused concern in the olive industry in the region. This spread may result from use of Verticillium dahliae-infected planting material, which can extend distribution of the highly virulent, defoliating (D) pathotype of V. dahliae to new areas. In this study, a molecular diagnostic method for the early in planta detection of D V. dahliae was developed, aimed especially at nursery-produced olive plants. For this purpose, new primers for nested PCR were designed by sequencing a 992-bp RAPD marker of the D pathotype. The use of the specific primers and different nested-PCR protocols allowed the detection of V. dahliae pathotype D DNA in infected root and stem tissues of young olive plants. Detection of the pathogen was effective from the very earliest moments following inoculation of olive plants with a V. dahliae pathotype D conidia suspension as well as in inoculated, though symptomless, plants.     

Identification ofTomato Mottle Taino Begomovirus strains in Cuban potato fields

The presence of a begomovirus in potato plants with yellow mottle symptoms was determined for the first time in Cuba. The incidence of typical begomovirus-like symptoms in potato plants in some regions of Havana province (Güira de Melena, San José de las Lajas, Güines and Boyeros) during the growing seasons from 1992 to 1998 was in general low. However, in some cultivars belonging to the National Program for Potato Genetic Improvement, the incidence reached 100%. Yield losses, determined in 1992 and 1994, ranged as high as 19% to 56.33% depending on the cultivar. Characterization of the causal agent was done by light microscopy, host range (graft and mechanical transmission), DNA hybridizations, polymerase chain reaction, and restriction fragment length polymorphism analysis. Nucleotide sequence of the amplified fragments revealed the presence ofTomato mottle Taino virus. The virus was transmittedvia tubers and has been detected in mixed infections withPotato virus X and withPotato leaf roll virus. http://www.phytoparasitica.org posting Oct. 20, 2003. The first two authors contributed equally to this work.     

Kaolin treatment to reduce pomegranate sunburn

The aim of this study was to reduce pomegranate losses due to sunburn in south-western Spain. Summer temperatures may rise above 45 °C, resulting in sunburn damage to the fruits, which may inflict losses of up to 40% of the total yield. Data were obtained in two consecutive years. Surround^® WP, a product comprising processed, refined kaolin, was sprayed over the whole canopy and fruits four times at 2–3-week intervals from mid-June to early August, the first application at 5% and the other three at 2.5%. The resulting white coating significantly reduced fruit and leaf surface temperatures relative to the control by averages of 4.9 and 2.5 °C, respectively. Sunburn damage of fruits was reduced from 21.9% in untreated control to 9.4% in the Surround WP-treated fruits.